改良硬膜外阻滞结合手法正脊治疗腰椎间盘突出症

目的:总结改良硬膜外阻滞结合手法正脊治疗腰椎间盘突出症的临床体会。方法:应用改良硬膜外阻滞结合手法正脊治疗156例腰椎间盘突出症患者。每10天1次,3次1个疗程。结果:优82例,良44例,可18例,差12例。优良率:80.7%;总有效率:92.3%。结论:改良硬膜外阻滞结合手法正脊治疗腰椎间盘突出症简单,安全,实用,疗效明显。     

结核病IL-10、IL-12、IL-18、TNF-α及TGF-β1免疫机制的研究

目的探讨IL-12、IL-18、IL-10和TNF-α与TGF-β1在结核病免疫发病机制中的作用。方法用双抗体夹心ELISA及ABC-ELISA法检测49例结核性胸腔积液患者胸水中和血清中以上5种细胞因子的水平,并进行相关性分析。结果从总体来看,除IL-18外,IL-12、TNF-α、IL-10胸水中含量都远远超过血清中的含量,而TGF-β1血清中含量远远超过胸水中含量(P<0.05或0.01)。结论 IL-18可能用作加强疫苗保护性的一种辅助因子;某种细胞因子不是对所有患者都有效,其作用与体内细胞因子间的平衡有关。     

Aerosol Delivery of Nanoparticles in Uniform Mannitol Carriers Formulated by Ultrasonic Spray Freeze Drying

Purpose

While most examples of nanoparticle therapeutics have involved parenteral or IV administration, pulmonary delivery is an attractive alternative, especially to target and treat local infections and diseases of the lungs. We describe a successful dry powder formulation which is capable of delivering nanoparticles to the lungs with good aerosolization properties, high loadings of nanoparticles, and limited irreversible aggregation.

Methods

Aerosolizable mannitol carrier particles that encapsulate nanoparticles with dense PEG coatings were prepared by a combination of ultrasonic atomization and spray freeze drying. This process was contrasted to particle formation by conventional spray drying.

Results

Spray freeze drying a solution of nanoparticles and mannitol (2 wt% solids) resulted in particles with an average diameter of 21?±?1.7 μm, regardless of the fraction of nanoparticles loaded (0–50% of total solids). Spray freeze dried (SFD) powders with a 50% nanoparticle loading had a fine particle fraction (FPF) of 60%. After formulation in a mannitol matrix, nanoparticles redispersed in water to < 1 μm with hand agitation and to < 250 nm with the aid of sonication. Powder production by spray drying was less successful, with low powder yields and extensive, irreversible aggregation of nanoparticles evident upon rehydration.

Conclusions

This study reveals the unique advantages of processing by ultrasonic spray freeze drying to produce aerosol dry powders with controlled properties for the delivery of therapeutic nanoparticles to the lungs.     

Development of hemiasterlin derivatives as potential anticancer agents that inhibit tubulin polymerization and synergize with a stilbene tubulin inhibitor

Hemiasterlins are cytotoxic tripeptides with antimicrotubule activity originally isolated from marine sponges. We have developed new hemiasterlin derivatives BF65 and BF78 that are highly potent to induce cancer cell death in the low nanomolar range. Examination of their mechanisms of cell cycle arrest and disruption of microtubules revealed an unusual characteristic in addition to anti-tubulin effect. Immunofluorescence staining revealed that A549 lung carcinoma cells treated with BF65 or BF78 exhibited both monopolar and multipolar mitotic spindles. Centrosomes were separated with short spindle microtubules in cells with multipolar spindles. In vitro tubulin polymerization assay confirmed that both BF65 and BF78 were highly potent to inhibit tubulin polymerization. These two compounds induced the formation of monoastral spindles suggesting that they might be inhibitors of mitotic kinesins such as KSP/Eg5. However, kinetic measurement of microtubule activated kinesin ATPase activity demonstrated that unlike the positive control monastrol, neither BF65 nor BF78 suppressed KSP/Eg5 activity. Hence the effect may be a variant form of tubulin inhibition. Similar to vinca alkaloids, BF compounds synergized with a colchicine site microtubule inhibitor stilbene 5c both in vitro and in vivo, which may provide a potential drug combination in the future clinical application.     

药检室在医院药学中的职能及功能拓展

目的 拓展医院药检室的职能.方法 综合分析探讨医院药检室的主要职能和功能拓展.结果 药检室的职能正从对自制制剂的质量控制扩大到对全院药品质量监督,从与各临床科室交流制剂问题到临床药学,从单一化工作到开展新制剂、新剂型等方面的科研研究.结论 药检室应根据实际情况,制订发展计划,在医院药学服务中发挥作用.     

Preparation and in vivo evaluation of immediate-release pellet containing celecoxib solid dispersion

The aim of this study was to make use of small size of immediate-release (IR) pellet and amorphous state of solid dispersion to increase solubility of celecoxib (CLX), a drug in BCS class II. Primary, binary and ternary solid dispersions were developed to choose the final components for solid dispersion. A ternary novel solid dispersion was prepared by incorporation of one aqueous soluble polymer (povidone k17; PVP 17PF), Methacrylate copolymer-based gastric soluble polymer (Eudragit^? EPO) and one pH modulator (MgO). This combination was effective to increase solubility in pH 1.2 up to 25?C30?%. The mechanism of solubility enhancement was proven by DSC, PRXD, and FT-IR. Accordingly, hydrogen bonding or electrostatic interaction of CLX with PVP/Eudragit^? EPO was the main cause to form the amorphous state of CLX within polymer cluster which increasing solubility of drug. Besides, MgO played an important role to change microenviroment for solid dispersion. Pellets containing this solid dispersion were prepared by extrusion and spheronization technique. Effect of four kinds of additive (calcium hydrogen phosphate dihydrate, NaHCO₃, crospovidone, and sodium dodecyl sulfate) on dissolution of CLX from IR pellet was also determined. Because of highest dissolution rate, formulation using sodium dodecyl sulfate was used for pharmacokinetics study. Solid dispersion-IR pellet formulation presented bioequivalence and lower variability in comparison with reference product.     

The role of MAPK and Nrf2 pathways in ketanserin-elicited attenuation of cigarette smoke-induced IL-8 production in human bronchial epithelial cells

Cigarette smoking is a major risk factor in chronic obstructive pulmonary disease (COPD) with chronic airway inflammation as a key feature. Blockade of serotonin receptor 2A (5-HTR(2A)) with ketanserin has been found to improve lung function in COPD patients. Furthermore, ketanserin has been shown to possess anti-inflammatory properties in vivo. In this study, we investigated the antioxidative and anti-inflammatory properties of ketanserin and its underlying mechanism of action on cigarette smoke-induced interleukin (IL)-8 release in vitro. Primary normal human bronchial epithelial cells and human bronchial epithelial cell line (BEAS-2B) were treated with or without ketanserin prior to exposure to cigarette smoke medium (CSM). Exposure to CSM caused elevation of both mRNA and release of IL-8 with increased phosphorylation of p38 and extracellular signal-regulated kinases 1 and 2 (ERK1/2). Consistently, CSM-induced IL-8 release was blocked by SB203580, U0126, or MEK1 small interfering RNA (siRNA) but not SP600125. On the other hand, CSM caused a dose-dependent decrease in the ratio of reduced glutathione to oxidized glutathione (rGSH/GSSG) together with an increased translocation of Nrf2 to the nucleus demonstrated by Western blot analysis. Knock down of Nrf2 by siRNA completely blocked CSM-induced IL-8 release. Ketanserin suppressed CSM-induced IL-8 release by inhibiting p38, ERK1/2 MAPK, and Nrf2 signaling pathways and partially inhibited CSM-induced reduction of rGSH/GSSG ratio. Our data demonstrated the novel antioxidative and anti-inflammatory role of ketanserin via the Nrf2 signaling pathway in CSM-exposed human bronchial epithelial cells. This may open up new perspectives in the development of novel therapeutic targets in the treatment of cigarette smoke-related COPD.     

Characterizing the effects of Eugenol on neuronal ionic currents and hyperexcitability

Rationale

Eugenol (EUG, 4-allyl-2-methoxyphenol), the main component of essential oil extracted from cloves, has various uses in medicine because of its potential to modulate neuronal excitability. However, its effects on the ionic mechanisms remains incompletely understood.

Objectives

We aimed to investigate EUG’s effects on neuronal ionic currents and excitability, especially on voltage-gated ion currents, and to verify the effects on a hyperexcitability-temporal lobe seizure model.

Methods

With the aid of patch-clamp technology, we first investigated the effects of EUG on ionic currents in NG108-15 neuronal cells differentiated with cyclic AMP. We then used modified Pinsky–Rinzel simulation modeling to evaluate its effects on spontaneous action potentials (APs). Finally, we investigated its effects on pilocarpine-induced seizures in rats.

Results

EUG depressed the transient and late components of I _Na in the neurons. It not only increased the degree of I _Na inactivation, but specifically suppressed the non-inactivating I _Na (I _Na(NI)). Its inhibition of I _Na(NI) was reversed by tefluthrin. In addition, EUG diminished L-type Ca²⁺ current and delayed rectifier K⁺ current only at higher concentrations. EUG’s effects on APs frequency reduction was verified by the simulation modeling. In pilocarpine-induced seizures, the EUG-treated rats showed no shorter seizure latency but a lower seizure severity and mortality than the control rats. The EUG’s effect on seizure severity was occluded by the I _Na(NI) antagonist riluzole.

Conclusion

The synergistic blocking effects of I _Na and I _Na(NI) contributes to the main mechanism through which EUG affects the firing of neuronal APs and modulate neuronal hyperexcitability such as pilocarpine-induced temporal lobe seizures.     

High-performance liquid chromatographic analysis for quantitation of marker compounds of Artemisia capillaris thunb.

Two stable high-performance liquid chromatography (HPLC) methods were developed that could quantitatively analyze 10 major marker compounds of Artemisia capillaris Thunb and could also distinguish among ‘Injinho’ and ‘Myeon-injin’ and ‘Haninjin’ — A. capillaris collected in autumn, A. capillaris collected in spring and A. iwayomogi, which can be misused as ‘Injinho’ in Korean herbal drug markets. The first HPLC method was a reversed-phase chromatography using a C18 column with an isocratic solvent system of phosphoric acid (0.05%) and acetonitrile at the flow rate of 1.0 mL/min, ultraviolet (UV) detection wavelength at 254 nm and column temperature at 40°C. Calibration and quantitation were made by using acetaminophen as an internal standard (I.S-A) and chlorogenic acid (1) was determined within 20 min. The second HPLC method was a reversed-phase chromatography using a C18 column with a gradient solvent system of phosphate buffer (0.015 M, pH 6) and acetonitrile at the flow rate of 1.0 mL/min, UV detection wavelength at 254 nm and column temperature at 40°C. Calibration and quantitation were made by using ethylparaben as an internal standard (I.S-B) and 3,5-di-O-caffeoylquinic acid (2), 3,4-di-O-caffeoylquinic acid (3), 4,5-di-O-caffeoylquinic acid (4), hyperoside (5), isoquercitrin (6), isorhamnetin 3-O-robinobioside (7), isorhamnetin-3-O-galactoside (8), isorhamnetin-3-O-glucoside (9) and scoparone (10) were determined within 60 min. Pattern recognition analysis of data from the 60 samples classified them clearly into three groups. These assay methods could be applied for QA/QC of A. capillaris and Artemisia iwayomogi.     

The mechanism of carvacrol-evoked [Ca2+]i rises and non-Ca2+-triggered cell death in OC2 human oral cancer cells

Carvacrol is one of the main substances of essential oil which triggers intracellular Ca²⁺ mobilization and causes cytotoxicity in diverse cell models. However, the mechanism of carvacrol-induced Ca²⁺ movement and cytotoxicity is not fully understood. This study examined the effect of carvacrol on cytosolic free Ca²⁺ concentrations ([Ca²⁺]_i), cell viability and apoptosis in OC2 human oral cancer cells. Carvacrol induced a [Ca²⁺]_i rise and the signal was reduced by removal of extracellular Ca²⁺. Carvacrol-induced Ca²⁺ entry was not altered by store-operated Ca²⁺ channel inhibitors and protein kinase C (PKC) activator, but was inhibited by a PKC inhibitor. In Ca²⁺ -free medium, treatment with the endoplasmic reticulum Ca²⁺ pump inhibitor thapsigargin (TG) or 2,5-di-tert-butylhydroquinone (BHQ) inhibited carvacrol-induced [Ca²⁺]_i rise. Conversely, incubation with carvacrol inhibited TG or BHQ-induced [Ca²⁺]_i rise. Inhibition of phospholipase C (PLC) with U73122 abolished carvacrol-induced [Ca²⁺]_i rise. Carvacrol decreased cell viability, which was not reversed when cytosolic Ca²⁺ was chelated with BAPTA-AM (1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid-acetoxymethyl ester). Carvacrol-induced apoptosis and activation of reactive oxygen species (ROS) and caspase-3. Together, carvacrol induced a [Ca²⁺]_i rise by inducing PLC-dependent Ca²⁺ release from the endoplasmic reticulum and Ca²⁺ entry via PKC-sensitive, non store-operated Ca²⁺ channels. Carvacrol-induced ROS- and caspase-3-associated apoptosis.